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HEK-293T cell line adaptation for suspension and serum free culture medium for large-scale production of lentiviral vectors

Grant number: 17/21737-2
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): February 01, 2018
Effective date (End): January 31, 2020
Field of knowledge:Health Sciences - Pharmacy
Principal Investigator:Kamilla Swiech Antonietto
Grantee:Ana Luiza Oliveira Lomba
Home Institution: Hemocentro de Ribeirão Preto. Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto da USP (HCMRP). Secretaria da Saúde (São Paulo - Estado). Ribeirão Preto , SP, Brazil
Associated research grant:13/08135-2 - CTC - Center for Cell-Based Therapy, AP.CEPID

Abstract

The use of chimeric antigen receptor (CAR) modified T lymphocyte immunotherapy has been shown to be effective in the treatment of chemotherapy and / or relapse resistant leukemias and lymphomas. Currently, lentiviral vectors are considered the most used gene modification vehicle of this therapy, offering significant advantages. Thus, with the growing interest in these vectors and the high demand, it is necessary to develop scalable, reproducible and high-quality production processes of large vector stock volumes. In this context, the main objective of this project is to adapt the HEK-293T cell line for suspension cultures by using serum-free culture media in order to establish a platform for the production of lentiviral vectors that allows future scale-up. The adaptation to serum free media and selection of the most efficient culture medium in terms of cell growth and vector production will be performed in static systems. The spinner flask (stirred conditions) will be used to adapt the cell to the suspension culture. After cell adaptation to the suspension culture, cultures will be carried out also in spinner flasks, to characterize the cell growth kinetics and the production of vectors through the transient transfection. Kinetics of cell growth and lentivirus production will be compared to those obtained in adherent cultures in culture medium containing fetal bovine serum (DMEM + 10% FBS). We hope to develop an efficient, rapid and reproducible platform for the production of lentiviral vectors that will contribute significantly to immunotherapy research. (AU)

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